Macrolide compounds

ABSTRACT

Compounds of formula (1) ##STR1## wherein R 1  represents a methyl, ethyl or isopropyl group; Y 1  is --CH 2  --, Y 2  is --CH-- and X represents ##STR2## [where R 2  represents a hydrogen atom or a group OR 6  (where OR 6  is a hydroxy group or a substituted hydroxyl group having up to 25 carbon atoms) and R 3  represents a hydrogen atom, or R 2  and R 3  together with the carbon atom to which they are attached represent &gt;C═O, &gt;C═CH 2  or &gt;C═NOR 7  (where R 7  represents a hydrogen atom, a C 1-8  alkyl group or a C 3-8  alkenyl group and the group &gt;C═NOR 7  is in the E configuration)] or --Y 1  --X--Y 2  represents --CH═CH--CH-- or --CH 2  --CH═C--; R 4  represents a group OR 6  as defined above and R 5  represents a hydrogen atom, or R 4  and R 5  together with the carbon atom to which they are attached represent &gt;C═O or &gt;C═NOR 7a  ( where R 7a  is as defined above for R 7 ); and one of R 8  and R 9  represents an alkoxyalkoxy group optionally interrupted by an oxygen atom or an alkoxy group and the other represents a hydrogen atom or R 8  and R 9  together with the carbon atom to which they are attached represent &gt;C═NOR 7b  (where R 7b  is as defined above for R 7 ), and salts thereof. 
     These compounds may be used to control nematode, acarine, insect or other pests.

This application is a continuation of application Ser. No. 07/350,333,filed May 9, 1989, now abandoned.

This invention relates to novel macrolide compounds, to processes fortheir preparation and to compositions containing them.

Thus, in one aspect, the invention particularly provides the compoundsof formula (I): ##STR3## wherein R¹ represents a methyl, ethyl orisopropyl group;

Y¹ is --CH₂ --, Y² is --CH-- and X represents ##STR4## [where R²represents a hydrogen atom or a group OR⁶ (where OR⁶ is a hydroxy groupor a substituted hydroxyl group having up to 25 carbon atoms) and R³represents a hydrogen atom, or R² and R³ together with the carbon atomto which they are attached represent >C═O, >C═CH₂ or >C═NOR⁷ (where R⁷represents a hydrogen atom, a C₁₋₈ alkyl group or a C₃₋₈ alkenyl groupand the group >C═NOR⁷ is in the E configuration)] or --Y¹ --X--Y² --represents --CH═CH--CH-- or --CH₂ --CH═C--;

R⁴ represents a group OR⁶ as defined above and R⁵ represents a hydrogenatom, or R⁴ and R⁵ together with the carbon atom to which they areattached represent >C═O or >C═NOR^(7a) (where R^(7a) is as defined abovefor R⁷); and

one of R⁸ and R⁹ represents an alkoxyalkoxy group optionally interruptedby an oxygen atom or an alkoxy group and the other represents a hydrogenatom or R⁸ and R⁹ together with the carbon atom to which they areattached represent >C═NOR^(7b) (where R^(7b) is as defined above forR⁷), and salts thereof.

Compounds of formula (I) are of use as antibiotics. The compounds of theinvention are also useful as intermediates in the preparation of furtheractive compounds. When the compounds of formula (I) are to be used asintermediates, the group --OR⁶ when present will often be a protectedhydroxy group.

When R⁸ or R⁹ represents an alkoxyalkoxy group optionally interrupted byan oxygen atom or an alkoxy group it may be for example a C₁₋₆ alkoxyC₁₋₆ alkoxy group optionally interrupted by an oxygen atom or a C₁₋₆alkoxy group.

The group R⁶ when present in compounds of formula (I) may represent anacyl group e.g. a group of the formula R¹⁰ CO-- or R¹⁰ OCO-- (where R¹⁰is an aliphatic, araliphatic or aromatic group, for example an alkyl,alkenyl, alkynyl, cycloalkyl, aralkyl or aryl group), a formyl group, agroup R¹¹ which is as defined above for R¹⁰, a group R¹² SO₂ -- (whereR¹² is a C₁₋₄ alkyl or C₆₋₁₀ aryl group), a silyl group, a cyclic oracyclic acetal group, a group --CO(CH₂)_(n) CO₂ R¹³ (where R¹³ is ahydrogen atom or a group as defined above for R¹⁰ and n represents zero,1 or 2) or a group R¹⁴ R¹⁵ NCO-- (where R¹⁴ and R¹⁵ may eachindependently represent a hydrogen atom or a C₁₋₄ alkyl group).

Where R¹⁰ or R¹¹ are alkyl groups, they may be for example C₁₋₈ alkylgroups, e.g. methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,t-butyl or n-heptyl, which alkyl groups may also be substituted. WhereR¹⁰ is a substituted alkyl group it may be substituted by, for example,one or more halogen atoms (e.g. chlorine or bromine atoms), or acarboxy, C₁₋₄ alkoxy (e.g. methoxy, ethoxy), phenoxy or silyloxy group.Where R¹¹ is a substituted alkyl group it may be substituted by acycloalkyl e.g. cyclopropyl group.

Where R¹⁰ and R¹¹ are alkenyl or alkynyl groups, they preferably have2-8 carbon atoms and where R¹⁰ and R¹¹ are cycloalkyl groups, they maybe for example C₃₋₁₂ cycloalkyl, such as C₃₋₇ cycloalkyl, e.g.cyclopentyl groups.

Where R¹⁰ and R¹¹ are aralkyl groups, they preferably have 1-6 carbonatoms in the alkyl moiety, and the aryl group(s) may be carbocyclic orheterocyclic and preferably contain 4-15 carbon atoms e.g. phenyl.Examples of such groups include phenC₁₋₆ alkyl e.g. benzyl groups.

Where R¹⁰ and R¹¹ are aryl groups, they may be carbocyclic orheterocyclic and preferably have 4-15 carbon atoms e.g. phenyl.

When R⁶ is a group R¹² SO₂ --, it may be for example a methylsulphonylor p-toluenesulphonyl group.

Where R⁶ represents a cyclic acetal group, it may for example have 5-7ring members as in the tetrahydropyranyl group.

When R⁶ represents a silyl group or R¹⁰ contains a silyloxy substituent,the silyl moiety may carry three groups, which may be the same ordifferent, selected from alkyl, alkenyl, alkoxy, cycloalkyl, aralkyl,aryl and aryloxy groups. Such groups may be as defined above for R⁶ andparticularly include methyl, t-butyl and phenyl groups. Particularexamples at such silyloxy groups are trimethylsilyloxy andt-butyldimethylsilyloxy.

Where R⁶ represents a group --CO(CH₂)_(n) CO₂ R¹³, it may for example bea group --COCO₂ R¹³ or --COCH₂ CH₂ CO₂ R¹³ where R¹³ represents ahydrogen atom or a C₁₋₄ alkyl group (e.g. methyl or ethyl).

When R⁶ represents a group R¹⁴ R¹⁵ NCO--, R¹⁴ and R¹⁵ for example mayeach independently be a hydrogen atom or a methyl or ethyl group.

Where R⁷ or R^(7a) or R^(7b) represents a C₁₋₈ alkyl group, it may befor example a methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl ort-butyl group, and is preferably a methyl group.

When R⁷ or R^(7a) represents a C₃₋₈ alkenyl group it may be for examplean allyl group.

Where R⁸ or R⁹ represents a C₁₋₆ alkoxy group, it may be for example amethoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy or t-butoxygroup, and is preferably a methoxy group.

Where R⁸ or R⁹ represents a C₁₋₆ alkoxy C₁₋₆ alkoxy group optionallyinterrupted by an oxygen atom, it may be for example a methoxy C₁₋₆alkoxy group optionally interrupted by an oxygen atom, and is preferablya group --OCH₂ OCH₂ CH₂ OCH₃.

Compounds of formula (I) containing an acidic group may form salts withbases. Examples of such salts include alkali metal salts such as sodiumand potassium salts.

In the compounds of formula (I) R¹ preferably represents an isopropylgroup.

An important group of compounds of formula (I) is that in which Y¹ is--CH₂ --, Y² is --CH--, and X represents ##STR5##

A further important group of compounds of formula (I) is that in whichR⁴ is a hydroxy, methoxy or acyloxy (e.g. acetyloxy) group or R⁴ and R⁵together with the carbon atom to which they are attached represent>C═NOCH₃. R⁴ preferably represents a hydroxyl group.

In a particular preference, R¹ in the compounds of formula (I) is anisopropyl group, Y¹ is --CH₂ --, Y² is --CH--, X is ##STR6## in which R²is a hydroxy, ethoxy or acetyloxy group and R³ is hydrogen atom, or R²and R³ together with the carbon atom to which they are attachedrepresents >C═O, >C═CH₂ or >C═NOCH₃ (where the group >C═NOCH₃ is in theE configuration), or R² and R³ each represents a hydrogen atom; R⁴ is ahydroxy or acetoxy group and R⁵ is a hydrogen atom; and one of R⁸ and R⁹represents a methoxy group or a group --OCH₂ OCH₂ CH₂ OCH₃ and the otherrepresents a hydrogen atom or R⁸ and R⁹ together with the carbon atom towhich they are attached represent >C═NOCH₃.

Important active compounds according to the invention are those offormula (I) in which:

R¹ is an isopropyl group, Y¹ is --CH₂ --, Y² is --CH--, X represents--CH₂ --, R⁴ is a hydroxyl group, R⁵ is a hydrogen atom, R⁸ is a methoxygroup and R⁹ is a hydrogen atom;

R¹ is an isopropyl group, Y¹ is --CH₂ --, Y² is --CH--, X represents>C═NOCH₃, R⁴ is an acetoxy group, R⁵ is a hydrogen atom, R⁸ is ahydrogen atom and R⁹ is a methoxy group;

R¹ is an isopropyl group, Y¹ is --CH₂ --, Y² is --CH--, X represents>C═NOCH₃, R⁴ is an acetoxy group, R⁵ is a hydrogen atom, R⁸ is a methoxygroup and R⁹ is a hydrogen atom; and

R¹ is an isopropyl group, Y¹ is --CH₂ --, Y² is --CH--, X represents>C═NOCH₃, R⁴ is an acetoxy group, R⁵ is a hydrogen atom, R⁸ is a group--OCH₂ OCH₂ CH₂ OCH₃ and R⁹ is a hydrogen atom.

As indicated previously, compounds according to the invention may be ofuse as intermediates for the preparation of further active compounds.When the compounds of the invention are to be used as intermediates, theR⁴ group may serve as a protecting group. It will be appreciated thatsuch a protecting group should have the minimum of additionalfunctionality to avoid further sites of reaction and should beselectively removable. Examples of groups serving as hydroxyl protectinggroups are well known and are described, for example, in "ProtectiveGroups in Organic Synthesis" by Theodora W. Greene. (Wiley-Interscience,New York 1981) and "Protective Groups in Organic Chemistry" by J. F. W.McOmie (Plenum Press, London, 1973). Examples of suitable R⁴ protectinggroups include phenoxyacetyl, silyloxyacetyl, (e.g.trimethylsilyloxyacetyl and t-butyldimethylsilyloxyacetyl), and silylsuch as trimethylsilyl and t-butyldimethylsilyl. Compounds of theinvention containing such groups will primarily be of use asintermediates. Other groups, such as acetyl, may serve as protectinggroups, but may also be present in final active compounds.

Compounds of the invention have antibiotic activity e.g. antihelminthicactivity, for example against nematodes, and in particular,anti-endoparasitic and anti-ectoparasitic activity.

The antibiotic activity of the compounds of formula (I) may, forexample, be demonstrated by their activity against parasitic nematodessuch as Nematospiroides dubius.

Ectoparasites and endoparasites infect humans and a variety of animalsand are particularly prevalent in farm animals such as pigs, sheep,cattle, goats and poultry (e.g. chickens and turkeys), horses, rabbits,game-birds, caged birds, and domestic animals such as dogs, cats, guineapigs, gerbils and hamsters. Parasitic infection of livestock, leading toanaemia, malnutrition and weight loss is a major cause of economic lossthroughout the world.

Examples of genera of endoparasites infecting such animals and/or humansare Ancylostoma, Ascaridia, Ascaris, Aspicularis, Brugia, Bunostomum,Capillaria, Chabertia, Cooperia, Dictyocaulus, Dirofilaria, Dracunculus,Enterobius, Haemonchus, Heterakis, Loa, Necator, Nematodirus,Nematospiroides (Heligomoroides), Nippostrongylus, Oesophagostomum,Onchocerca, Ostertagia, Oxyuris, Parascaris, Strongylus, Strongyloides,Syphacia, Toxascaris, Toxocara, Trichonema, Trichostrongylus,Trichinella, Trichuris, Triodontophorus, Uncinaria and Wuchereria.

Examples of ectoparasites infecting animals and/or humans are arthropodectoparasites such as biting insects, blowfly, fleas, lice, mites,sucking insects, ticks and other dipterous pests.

Examples of genera of such ectoparasites infecting animals and/or humansare Ambylomma, Boophilus, Chorioptes, Culliphore, Demodex, Damalinia,Dermatobia, Gastrophilus, Haematobia, Haematopinus, Haemophysalis,Hyaloma, Hypoderma, Ixodes, Linognathus, Lucilia, Melophagus, Oestrus,Otobius, Otodectes, Psorergates, Psoroptes, Rhipicephalus, Sarcoptes,Stomoxys and Tabanus.

Furthermore, the compounds of formula (I) are also of use in combatinginsect, acarine and nematode pests in agriculture, horticulture,forestry, public health and stored products. Pests of soil and plantcrops, including cereals (e.g. wheat, barley, maize and rice) vegetables(e.g. soya), fruit (e.g. apples, vines and citrus) as well as root crops(e.g. sugarbeet, potatoes) may usefully be treated. Particular examplesof such pests are fruit mites and aphids such as Aphis fabae,Aulacorthum circumflexum, Myzus persicae, Uephotettix cincticeps,Nilparvata lugens, Panonychus ulmi, Phorodon humuli, Phyllocoptrutaoleivora, Tetranychus urticae and members of the genera Trialeuroides;nematodes such as members of the genera Aphelencoides, Globodera,Heterodera, Meloidogyne and Panagrellus; lepidoptera such as Heliothis,Plutella and Spodoptera; grain weevils such as Anthonomus grandis andSitophilus granarius; flour beetles such as Tribolium castaneum; fliessuch as Musca domestica; fire ants; leaf miners; Pear psylla; Thripstabaci; cockroaches such as Blatella germanica and Periplaneta americanaand mosquitoes such as Aedes aegypti.

According to the invention we therefore provide the compounds of formula(I) as defined above, which may be used as antibiotics. In particular,they may be used in the treatment of animals and humans withendoparasitic, ectoparasitic and/or fungal infections and inagriculture, horticulture, or forestry as pesticides to combat insect,acarine and nematode pests. They may also be used generally aspesticides to combat or control pests in other circumstances, e.g. instores, buildings or other public places or location of the pests. Ingeneral the compounds may be applied either to the host (animal or humanor plants or other vegetation) or to the pests themselves or a locusthereof.

The compounds of the invention may be formulated for administration inany convenient way for use in veterinary or human medicine and theinvention therefore includes within its scope pharmaceuticalcompositions comprising a compound in accordance with the inventionadapted for use in veterinary or human medicine. Such compositions maybe presented for use in conventional manner with the aid of one or moresuitable carriers or excipients. The compositions of the inventioninclude those in a form especially formulated for parenteral (includingintramammary administration), oral, rectal, topical, implant,ophthalmic, nasal or genito-urinary use.

The compounds of formula (I) may be formulated for use in veterinary orhuman medicine according to the general methods described in UK PatentSpecification 2166436.

The total daily dosages of the compounds of the invention employed inboth veterinary and human medicine will suitably be in the range 1-2000μg/kg bodyweight, preferably from 50-1000 μg/kg and these may be givenin divided doses, e.g. 1-4 times per day.

The compounds according to the invention may be formulated in anyconvenient way for horticultural or agricultural use and the inventiontherefore includes within its scope compositions comprising a compoundaccording to the invention adapted for horticultural or agriculturaluse. Such formulations include dry or liquid types, for example dusts,including dust bases or concentrates, powders, including soluble orwettable powders, granulates, including microgranules and dispersiblegranules, pellets, flowables, emulsions such as dilute emulsions oremulsifiable concentrates, dips such as root dips and seed dips, seeddressings, seed pellets, oil concentrates, oil solutions, injectionse.g. stem injections, sprays, smokes and mists.

Generally such formulations will include the compound in associationwith a suitable carrier or diluent. Such carriers and dilutents are asdescribed in UK Patent Specification 2166436.

In the formulations, the concentration of active material is generallyfrom 0.01 to 99% and more preferably between 0.01% and 40% by weight.

Commercial products are generally provided as concentrated compositionsto be diluted to an appropriate concentration, for example from 0.001 to0.0001% by weight, for use.

The rate at which a compound is applied depends upon a number of factorsincluding the type of pest involved and the degree of infestation.However, in general, an application rate of 10 g/ha to 10 kg/ha will besuitable; preferably from 10 g/ha to 1 kg/ha for control of mites andinsects and form 50 g/ha to 10 kg/ha for control of nematodes.

The antibiotic compounds of the invention may be administered or used incombination with other active ingredients.

The compounds according to the invention may be prepared by a number ofprocesses as described in the following where R¹ -R⁹, X, Y¹ and Y² areas defined for general formula (I) unless specified otherwise. In someof these processes it may be necessary to protect a hydroxyl group atthe 5-, 13- and/or 23-position in the starting material prior toeffecting the reaction described. In such cases it may then be necessaryto deprotect the same hydroxyl group once the reaction has occurred toobtain the desired compound of the invention. Conventional methods ofprotection and deprotection may be used, for example as described in theaforementioned books by Greene and McOmie.

According to one process (A), a compound of formula (I) in which R⁸ andR⁹ together with the carbon atom to which they are attached represent>C═NOR^(7b) (where R^(7b) is an previously defined) may be prepared froma compound of formula (II) ##STR7## (wherein R⁴ represents a group OR⁶where OR⁶ is as previously defined or represents a protected hydroxylgroup e.g. acetoxy) by reaction with a reagent H₂ NOR^(7b) (where R^(7b)is as previously defined) or a salt thereof followed, if necessary, byremoval of any protecting groups present.

The oximation reaction may conveniently be effected at a temperature inthe range -20° to +100° C., e.g. -10° to +50° C. It is convenient to usethe reagent H₂ NOR^(7b) in the form of a salt, for example an acidaddition salt such as the hydrochloride. When such a salt is employedthe reaction may be carried out in the presence of an acid bindingagent.

Solvents which may be employed include alcohols (e.g. methanol orethanol), amides (e.g. N,N-dimethylformamide, N,N-dimethylacetamide orhexamethylphosphoramide), ethers (e.g. cyclic cyclic ethers such astetrahydrofuran or dioxan, and acylic ethers such as dimethoxyethane ordiethylether), nitriles (e.g. acetonitrile), sulphones (e.g. sulpholane)and hydrocarbons such as halogenated hydrocarbons (e.g. methylenechloride), as well as mixtures of two or more such solvents. Water mayalso be employed as a cosolvent.

When aqueous conditions are employed the reaction may conveniently bebuffered with an appropriate acid, base or buffer.

Suitable acids include mineral acids, such as hydrochloric or sulphuricacid, and carboxylic acid such as acetic acid. Suitable bases includealkali metal carbonates and bicarbonates such as sodium bicarbonate,hydroxides such as sodium hydroxide, and alkali metal carboxylates suchas sodium acetate. A suitable buffer is sodium acetate/acetic acid.

According to another process (B), a compound of formula (I) in which R⁸or R⁹ represents an alkoxyalkoxy group optionally interrupted by anoxygen atom or an alkoxy group may be prepared from a compound offormula (III) ##STR8## (wherein one of R⁸ and R⁹ represents a hydroxylgroup and the other represents a hydrogen atom and R⁴ represents a groupOR⁶ where OR⁶ is as previously defined or represents a protectedhydroxyl group e.g. acetoxy) by reaction with an etherifying agent,followed, if necessary, by removal of any protecting groups present.

Etherification may be effected using a reagent of formula R^(ay) (whereR^(a) is an alkyl group or an alkoxyalkyl group optionally interruptedby an oxygen atom and Y represents a leaving group such a a halogen atom(e.g. chlorine, bromine or iodine) or a hydrocarbylsulphonyloxy group(e.g. mesyloxy or tosyloxy) or a haloalkanoyloxy group (e.g.dichloroacetoxy). When the etherification reaction is carried out usinga halide it is preferable that a suitable base such as an amine (e.g.diisopropylethylamine) is also present.

Etherification may also be effected using a trialkyloxonium salt (e.g. atrialkyloxonium tetrafluoroborate salt), preferably in the presence of asuitable base e.g. 1,8-bis(dimethylamino)napthalene.

Solvents which may be employed in the above etherification reactionsinclude ethers such as diethyl ether and hydrocarbons such ashalogenated hydrocarbons (e.g. dichloromethane). The reaction mayconveniently be carried out at a temperature in the range of 0° to 50°C., preferably at room temperature.

In yet another process (C), a compound of formula (I) in which R⁴ is ahydroxyl group may be prepared from a corresponding compound of formula(I) in which R⁴ is a substituted hydroxyl group. The conversion willusually be carried out in the context of removing a protecting groupsuch as referred to above.

Thus, deprotection of the compounds of the invention in which R⁴represents a protected hydroxyl group can be effected by conventionalmethods, for example those extensively described in the aforementionedtextbooks of McOmie and Greene. Thus, for example, when R⁴ is an acyloxygroup such as an acetoxy group the acetyl group may be removed by basichydrolysis, e.g. using sodium or potassium hydroxide or ammonia in anaqueous alcohol such as methanol to yield the compound of formula (I) inwhich R⁴ is a hydroxyl group.

In a further process (D), the compounds of the invention in which OR⁶ isa substituted hydroxyl group may generally be prepared by reacting thecorresponding 5 and/or 23-hydroxy compound with reagents serving to forma substituted hydroxyl group, followed, if necessary, by removal of anyprotecting groups present.

The reaction will in general be an acylation, sulphonylation,etherification, silylation, or acetalation, and the reaction may beeffected according to the general methods described in UK PatentSpecification 2176182. It will be appreciated that the etherificationreaction may be carried out on a compound of formula (III) to provide inone step a compound of formula (I) in which OR⁶ is alkoxy group and oneof R⁸ and R⁹ is an alkoxy group and the other is a hydrogen atom.

In a further process (E), a compound of formula (I) in which X ##STR9##is and R² and R³ together with the carbon atom to which they areattached represent >C═O may be prepared by oxidising a correspondingcompound of formula (I) wherein R² is a hydroxyl group and R³ is ahydrogen atom, followed, if necessary, by removal of any protectinggroups present. The reaction may be effected with an oxidising agentserving to convert a secondary hydroxyl group to an oxo group, whereby acompound of formula (I) is produced.

Suitable oxidising agents include quinones in the presence of water,e.g. 2,3-dichloro-5,6-dicyano-1,4-benzoquinone or2,3,5,6-tetrachloro-1,4-benzoquinone; a chromium (VI) oxidising agent,e.g. pyridinium dichromate or chromium trioxide in pyridine; a manganese(IV) oxidising agent, e.g. manganese dioxide in dichloromethane; anN-halosuccinimide, e.g. N-chlorosuccinimide or N-bromosuccinimide; adialkylsulphoxide e.g. dimethylsulphoxide, in the presence of anactivating agent such as N,N'-dicyclohexylcarbodiimide or an acylhalide, e.g. oxalyl choride; or a pyridine-sulphur trioxide complex.

The reaction may conveniently be effected in a suitable solvent whichmay be selected from a ketone, e.g. acetone; an ether, e.g. diethylether, dioxan or tetrahydrofuran; a hydrocarbon, e.g. hexane; ahalogenated hydrocarbon e.g. chloroform or methylene chloride; or anester, e.g. ethyl acetate or a substituted amide e.g. dimethylformamide.Combinations of such solvents either alone or with water may also beused. The choice of solvent will depend on the oxidising agent used toeffect the conversion.

The reaction may be carried out at a temperature of from -80° C. to +50°C.

In another process (F), a compound of formula (I) in which X is >C═NOR⁷may be prepared from the corresponding 23-keto compound of formula (I)in which X is >C═O by reaction with a reagent H₂ NOR⁷ (where R⁷ is asdefined previously). The reaction is preferably carried out using aboutone equivalent of the reagent H₂ NOR⁷ and may conveniently be effectedusing the conditions described in process (A) above.

In a particular embodiment of this process, compounds of formula (I) inwhich X represents >C═NOR⁷ and R⁸ and R⁹ together with the carbon atomto which they are attached represent >C═NOR^(7b) may be prepared fromcompounds of formula (II) in which X represents >C═O using twoequivalents of the reagent H₂ NOR⁷ under the conditions described inprocess (A) above. It will be appreciated that in the preparation of13,23-bisoximes of formula (I) from corresponding 13,23-diketones thegroups >C═NOR⁷ and >C═NOR^(7b) will be equivalent.

In a further process (G), a compound of formula (I) in which X is agroup >C═CH₂ may be prepared by reaction of a corresponding compound offormula (I) in which X is >C═O with an appropriate Witting reagent e.g.a phosphorane of formula (R¹⁶)₃ P═CH₂ (where R¹⁶ is C₁₋₆ alkyl or aryl,e.g. monocyclic aryl such as phenyl). Suitable reaction solvents includeethers such as tetrahydrofuran or diethyl ether or a dipolar aproticsolvent such as dimethylsulphoxide. The reaction may be carried out atany suitable temperature e.g. at 0° C.

Intermediate compounds of formula (II) in which R⁴ is substitutedhydroxyl group may be prepared from a compound of formula (III) in whichR⁴ is a substituted hydroxyl group by oxidation. Suitable oxidisingagents for the conversion include dialkylsulphoxides, e.g.dimethylsulphoxide, in the presence of an activating agent such asN,N'-dicyclohexylcarbodiimide or an acyl halide e.g. oxalyl chloride.The reaction may conveniently be effected in a suitable solvent such asa halogenated hydrocarbon, e.g. dichloromethane, at a temperature in therange of -80° to +50° C.

Intermediate compounds of formula (II) in which R⁴ is a hydroxy groupmay be prepared from the corresponding compounds of formula (II) inwhich R⁴ is a substituted hydroxyl group using the methods describedabove for the preparation of compounds of formula (I) in which R⁴ is ahydroxyl group.

Intermediate compounds of formula (III) in which R⁸ is a hydroxyl groupand R⁹ is a hydrogen atom may be prepared by oxidising a compound offormula (IV) ##STR10##

The oxidation may be effected for example with an oxidising agent suchas selenium dioxide, preferably in the presence of an activator such asa peroxide, e.g. tert-butyl hydroperoxide. The reaction may convenientlybe effected in an inert solvent such as a halogenated hydrocarbon e.g.dichloromethane, an ester, e.g. ethyl acetate or an ether, e.g.tetrahydrofuran, at a temperature in the range of 0° to 50° C.,preferably at room temperature.

Alternatively, a compound of formula (IV) may be treated with anoxidising agent described above in formic acid at a temperature of from20° to 100° C. e.g. 60° C. to provide a compound of formula (V).##STR11## which, upon acid hydrolysis, e.g. using hydrochloric acidprovides a compound of formula (III).

Intermediate compounds of formula (III) in which R⁸ is a hydrogen atomand R⁹ is a hydroxyl group may be prepared by reducing a compound offormula (II).

The reduction may be effected for example using a reducing agent such asa borohydride, for example an alkali metal borohydride such as sodiumborohydride or a lithium alkoxyaluminium hydride such as lithiumtributoxyaluminium hydride.

The reaction involving a borohydride reducing agent takes place in thepresence of a solvent such as an alkanol e.g. isopropyl alcohol orisobutyl alcohol conveniently at a temperature in the range of -30° to+80° C. e.g. at 0° C. The reaction involving a lithium alkoxyaluminiumhydride takes place in the presence of a solvent such as an ether e.g.tetrahydrofuran or dioxan conveniently at a temperature in the range of-78° to 0° C. e.g. at -78° C.

Intermediate compounds of formula (IV) in which Y¹ is --CH₂ --, Y² is--CH-- and X represents ##STR12## (where R² is a hydrogen atom or agroup OR⁶ and R³ is a hydrogen atom or R² and R³ together with thecarbon atom to which they are attached represent >C═O), R⁴ is a groupOR⁶ and R⁵ is a hydrogen atom are known compounds described in UK PatentSpecifications 2166436 and 2176182.

Intermediate compounds of formula (IV) in which --Y¹ --X--Y² --represents --CH═CH--CH-- or CH₂ --CH═C--, R⁴ is a group OR⁶ and R⁵ is ahydrogen atom are known compounds described in European PatentSpecification 215654.

Intermediate compounds of formula (IV) in which Y¹ is --CH₂ --, Y² is--CH-- and X represents >C═CH₂ may be prepared by reaction of acorresponding compound of formula (IV) in which X is >C═O with anappropriate Witting reagent according to the method of process (G)above.

Intermediate compounds of formula (IV) in which Y¹ is --CH₂ --, Y² is--CH--, X represents >C═NOR⁷ (where R⁷ is as previously defined), R⁴ isa group OR⁶ and R⁵ is a hydrogen atom or R⁴ and R⁵ together with thecarbon atom to which they are attached represent >C═O, or intermediatesin which X represents a group ##STR13## (where R² is a hydrogen atom ora group OR⁶ and R³ is a hydrogen atom) or X represents >C═NOR⁷ or --Y¹--X--Y² -- represents --CH═CH--CH-- or --CH₂ --CH═C-- and R⁴ and R⁵together with the carbon atom to which they are attached represent>C═NOR^(7a) may be prepared from the corresponding 5 and/or 23-ketocompounds of formula (IV) by reaction with a reagent H₂ NOR⁷ using theoximation reaction conditions described above. It will be appreciatedthat in the preparation of a 5,23-bisoxime of formula (IV) from acorresponding 5,23-diketone the groups >C═NOR⁷ and >C═NOR^(7a) will beequivalent.

Intermediates of formula (IV) in which R⁴ and R⁵ together with thecarbon atom to which they are attached represent >C═O may be prepared byoxidation of the corresponding 5-hydroxy compounds in which R⁴ is ahydroxy group.

The reaction may be effected with an oxidising agent serving to convertan allylic secondary hydroxyl group to an oxo group, whereby a compoundof formula (IV) is produced.

Suitable oxidising agents include, for example, transition metal oxides,such as manganese dioxide, and atmospheric oxygen in the presence of asuitable catalyst such as a finely divided metal e.g. platinum.

The oxidising agent will generally be used in excess over thestoichiometric quantity.

The reaction may conveniently be effected in a suitable solvent whichmay be selected from a ketone, e.g. acetone; an ether, e.g. diethylether, dioxan or tetrahydrofuran; a hydrocarbon, e.g. hexane; ahalogenated hydrocarbon e.g. chloroform or methylene chloride; or anester, e.g. ethyl acetate. Combinations of such solvents either alone orwith water may also be used.

The reaction may be carried out at a temperature of from -50° C. to +50°C., preferably from 0° to 30° C.

It will be apprciated that the processes described above for preparingintermediate compounds of formula (IV) may also be utilised for thepreparation of corresponding compounds of formulae (I), (II) and (III)and the present invention extends to cover such processes.

Intermediate compounds of formula (V) are novel compounds and constitutea further aspect of the present invention.

The compounds of formula (V) may, in general, be prepared by oxidising acorresponding compound of formula (IV). The oxidation may be effectedfor example using an oxidising agent such as selenium dioxide in formicacid at a temperature in the range of 20° to 100° C., e.g. 60° C.

Intermediate compounds of formula (V) in which X represents the group>C═NOR⁷ may also be prepared from a corresponding compound of formula(V) in which X represents the group >C═O by reaction with a reagent H₂NOR⁷ using the method of process (F) described above.

The invention is further illustrated by the following Preparations andExamples wherein the compound of formula (IV) above in which R¹ isisopropyl, Y¹ is --CH₂ --, Y² is --CH--, X represents ##STR14## (whereR² is a hydroxyl group and R³ is a hydrogen atom), R⁴ is a hydroxylgroup and R⁵ is a hydrogen atom is referred to as `Factor A`. Compoundsaccording to the invention are named with respect to Factor A. Alltemperatures are in 0° C.

INTERMEDIATE 1 (13R)-Hydroxy-23-desoxy Factor A, 5-acetate

23-Desoxy Factor A, 5-acetate (4.79 g, Example 112 in UK PatentSpecification 2176182) was added to a stirred mixture of seleniumdioxide (416 mg) and t-butyl hydroperoxide (3M in dichloromethane; 5 ml)in dichloromethane (30 ml). After stirring at room temperature for 30 hthe reaction mixture was diluted with ethyl acetate (200 ml), washedwith water and brine, and dried (Na₂ SO₄). The solvent was evaporatedand the residue purified by chromatography (250 g silica gel, Merck9385). Elution with ethyl acetate: light petroleum (1:4→1:2) affordedthe title compound (560 mg) as a pale yellow foam; ν_(max) (CHBr₃) 3600,3460 (OH), 1732 (OAc), 1712 (CO₂ R), 993 cm⁻¹ (C-O); δ(CDCl₃) valuesinclude 0.69 (3H, t, J 5 Hz), 2.15 (3H,s), 3.32 (1H,m), 3.72 (1H, d, J10 Hz), 4.05 (1H, d, J 5 Hz), 5.52 (2H,m).

INTERMEDIATE 2 (13R)-Formyloxy-23-keto Factor A, 5-acetate

To a slurry of selenium dioxide (120 mg) in formic acid (1 ml) stirringat 60° was added a solution of 23-keto Factor A, 5-acetate (420 mg,Example 18 in UK Patent Specification 2176182) in formic acid (3 ml).The reaction mixture was left stirring at 60° for 6 mins then was pouredinto water (150 ml) and extracted with diethyl ether (4×50 ml). Theorganic phase was dried (MgSO₄) and solvent removed to give a brownsolid which was purified by medium pressure column chromatography onsilica (100 g Merck kieselgel 60; 230-400 mesh). Elution withdichloromethane:ethyl acetate (16:1) gave the title compound as a creamfoam (103 mg); νmax (CHBr₃) 3480 (OH) and 1714 cm⁻¹ (ester and ketone);δ(CDCl₃) includes 0.86 (d, 6 Hz,3H), 0.97 (d,6 Hz, 3H), 1.02 (d,6Hz,3H), 1.07 (d,6 Hz,3H), 1.76 (s,3H), 3.32 (m,1H), 2.16 (s,3H), 4.06(d,6 Hz,1H), 5.02 (d,10 Hz,1H), 5.53 (m,2H), 8.08 (s,1H).

INTERMEDIATE 3 (13R)-Formyloxy-23(E)-methoxyimino Factor A, 5-acetate

To a solution of Intermediate 2 (80 mg) in methanol (8 ml) was added asolution of methoxyamine hydrochloride (29 mg) and sodium acetate (33mg) in water (0.7 ml). The reaction mixture was left stirring at roomtemperature for 3 h, then was poured into ether (40 ml) and washed withwater. The organic phase was dried (MgSO₄) and solvent removed to givethe title compound as a cream foam (79 mg); δ(CDCl₃) includes 0.91 (d,6Hz,3H), 0.97 (d,6 Hz,3H), 1.02 (d,6 Hz,3H), 1.07 (d,6 Hz,3H), 1.76(s,3H), 2.16 (s,3H)m, 3.28 (d,15 Hz,1H), 1.91 (d,15 Hz,1H), 3.32 (m,1H),3.83 (s,3H), 4.06 (d,6 Hz,1H), 5.04 (d,10 Hz,1H), 5.54 (m,2 H), 8.09(s,1H).

INTERMEDIATE 4 (13R)-Hydroxy-23(E)-methoxyimino Factor A, 5-acetate

a) To a solution of Intermediate 3 (65 mg) in methanol (5 ml) was added2N hydrochloric acid (0.1 ml). The reaction mixture was left stirring atroom temperature for 4 h, then was poured into dichloromethane (60 ml)and washed with saturated sodium bicarbonate solution and water (40 mlof each). The organic phase was dried (MgSO₄) and solvent removed togive a foam (65 mg) which was purified by medium pressure columnchromatography on silica (30 g, Merck Kieselgel 60, 230-400 mesh).Elution with dichloromethane:ethyl acetate (4:1) gave the title compoundas a white foam (39 mg); [α]_(D) ²¹ +126° (C═0.22, CH₂ Cl₂). δ(CDCl₃)includes 0.92 (d,6 Hz,3H), 0.96 (d,6 Hz,3H), 1.05 (d,6 Hz,3H), 1.12 (d,6Hz,3H), 1.77 (s,3H), 2.17 (s,3 H), 3.29 (d,15 Hz,1H), 1.91 (d,15 Hz,1H),3.32 (m,1H), 3.70 (dd10,2 Hz,1H), 3.83 (s,3H), 4.04 (d,6 Hz,1H), 5.54(m,2H).

b) To a slurry of selenium dioxide (460 mg) in formic acid (6 ml)stirring at 60° was added a solution of 23-keto Factor A, 5-acetate(1.80 g) in formic acid (16 ml). The reaction mixture was left stirringat 60° for 6 min, then was poured into water (500 ml) and extracted withdiethyl ether (4×200 ml). The organic phase was dried (MgSO₄) andsolvent removed to give a brown foam (1.89 g).

To a solution of this foam (1.89 g) in methanol (180 ml) was added asolution of methoxyamine hydrochloride (676 mg) and sodium acetate (760mg) in water (16 ml). The reaction mixture was left stirring at roomtemperature for 3 h. Diethyl ether (700 ml) was added and the resultingmixture was washed with water. The organic phase was dried (MgSO₄) andthe solvent removed to give a brown solid (1.89 g).

To a solution of this brown solid (1.89 g) in methanol (140 ml) wasadded 2N hydrochloric acid (3 ml) and the reaction mixture left stirringat room temperature for 2 h. Dichloromethane (1000 ml) was added and thesolution was washed with saturated sodium bicarbonate solution, waterand brine. The organic phase was dried (MgSO₄) and the solvent removedto give a brown foam (1.81 g) which was purified by medium pressurecolumn chromatography (480 g, Merck Kieselgel 60, 230-400 mesh). Elutionwith dichloromethane:ethyl acetate (5:1) gave the title compound as awhite foam (462 mg). Nmr as described above.

INTERMEDIATE 5 13-Keto-23(E)-methoxyimino Factor A, 5-acetate

To a solution of oxalyl chloride (0.24 ml) in freshly distilleddichloromethane (3.6 ml) stirring at -60° under nitrogen was added asolution of dimethyl sulphoxide (0.4 ml) in freshly distilleddichloromethane (3.6 ml). The solution was cooled to -65° and after 5mins a solution of Intermediate 4 (770 mg) in dichloromethane (6 ml) wasadded. The cooling bath was allowed to warm to -60° then the reactionmixture left a further 30 min stirring at -60° to -50°. Triethylamine(1.5 ml) was added and the reaction mixture allowed to warm to roomtemperature. The reaction mixture was then poured into dichloromethane(100 ml) and the solvent removed under vacuum. Diethyl ether (60 ml) wasadded and the triethylamine salt filtered off. The ether was removedunder vacuum to give a foam (760 mg) which was purified by mediumpressure column chromatography on silica (180 g, Merck Kieselgel 60,230-400 mesh). Elution with dichloromethane:ethyl acetate (14:1) gavethe title compound as a beige foam (450 mg); δ(CDCl₃) includes 0.92 (d,6Hz,3H), 0.96 (d,6 Hz,3H), 1.01 (d,6 Hz,3H), 1.18 (d,6 Hz,3H), 1.76(s,3H), 1.80 (s,3H), 2.16 (s,3H), 3.31 (d,15 Hz, 1H), 1.93 (d,15 Hz,1H),3.39 (m,1H), 3.84 (s,3H), 4.08 (d,6 Hz,1H), 5.54 (m,2H), 6.22 (t,9Hz,1H).

INTERMEDIATE 6 (13S)-Hydroxy-23(E)-methoxyimino Factor A, 5-acetate

To a solution of Intermediate 5 (620 mg) in ethanol (25 ml) stirring at0° was added a solution of sodium borohydride (4.9 ml of a 0.2M solutionin ethanol). The reaction mixture was left stirring at 0° for 30 min,then was poured into ethyl acetate (400 ml) and washed with 2Nhydrochloric acid, saturated sodium bicarbonate solution water andbrine. The organic phase was dried (MgSO₄) and solvent removed to give abeige foam (605 mg) which was purified by medium pressure columnchromatography on silica (180 g, Merck Kieselgel 60, 230-400 mesh).Elution with dichloromethane:ethyl acetate (10:1) gave the titlecompound as a white foam (502 mg); δ(CDCl₃) includes 0.92 (d,6 Hz,3H),0.97 (d,6 Hz,3H), 1.05 (d,6 Hz,3H), 1.16 (d,6 Hz,3H), 1.76 (s,3H), 2.16(s,3H), 3.29 (d,15 Hz,1H), 1.91 (d,15 Hz,1H), 3.32 (m,1H), 3.84 (s,3H),4.00 (broad s,1H), 4.06 (d,6 Hz,1H), 5.53 (m,2H)

EXAMPLE 1 23-Desoxy-(13R)-methoxy Factor A, 5-acetate

A solution of Intermediate 1 (47.8 mg) in dichloromethane (2 ml) wastreated with trimethyloxonium tetrafluoroborate (108 mg) and1,8-bis(dimethylamino)naphthalene (156 mg) under an atmosphere ofnitrogen. After 22 h at room temperature, the reaction mixture wasdiluted with ethyl acetate (50 ml), washed with 2M hydrochloric acid,saturated sodium bicarbonate solution and brine, and dried (Na₂ SO₄).The solvent was evaporated and the residue purified by flashchromatography (15 g silica gel, Merck 9385). Elution with ethylacetate:petroleum ether (1:4) afforded the title compound as a paleyellow foam (32 mg); λ_(max) (EtOH) 245.2 nm (ε29,600); ν_(max) (CHBr₃)1732 (OAc, 1710 cm⁻¹ (CO₂ R); δ(CDCl₃) values include 0.69 (3H, d, J5Hz), 0.94 (3H, d, J6 Hz), 1.04 (3H, d, J6 Hz), 1.08 (3H, d, J6 Hz), 1.76(3H, s), 3.11 (1H, d, J10 Hz), 3.16 (3H, s), 3.32 (1H, m), 4.05 (1H, d,J6 Hz), 5.54 (2H, m).

EXAMPLE 2 23-Desoxy-(13 R)-methoxy Factor A

Aqueous sodium hydroxide (1M; 60 μl) was added to a solution of Example1 (27 mg) in methanol (1 ml) at 0°. After 1.5 h at 0°, the reactionmixture was diluted with ethyl acetate (50 ml), washed with water andbrine and dried (Na₂ SO₄). The solvent was evaporated and the residuepurified by flash chromatography (15 g silica gel, Merck 9385). Elutionwith ethyl acetate:light petroleum (1:3) afforded the title compound asa white foam (22.7 mg); λ_(max) (EtOH) 245 nm (ε28,400); ν_(max) (CHBr₃)3540 (OH), 1708 cm⁻¹ (CO₂ R); δ(CDCl₃) values include 0.68 (3H, d, J3Hz), 0.94 (3H, d, J6 Hz), 1.05 (3H, d, J6 Hz), 1.08 (3H, d, J6 Hz), 1.87(3H, s), 3.11 (1H, d, J10 Hz), 3.16 (3H, s), 3.28 (1H,m) 3.96 (1H, d, J5Hz), 4.29 (1H, t, 5 Hz).

EXAMPLE 3 (13R)-Methoxy-23(E)-methoxyimino Factor A,5-acetate

A mixture of Intermediate 4 (47 mg), trimethyloxonium tetrafluoroborate(106 mg) and 1,8-bis(dimethylamino)naphthalene (153 mg) indichloromethane (0.4 ml) was stirred in an atmosphere of nitrogen atroom temperature for 18 h. Ice-water (15 ml) was added and the mixturewas extracted with ether (2×25 ml). The extracts were washed with 5%sodium hydrogen carbonate and water and dried. Removal of solvent gave awhite gum which was purified by medium pressure column chromatography onsilica (40 g, Merck Kieselgel 60, 230-400 mesh). Elution withdichloromethane:ether (6:1) gave the title compound as a white foam(26.5 mg); [α]_(D) ²² +101° (C,0.4, CH₂ Cl₂); λmax (EtOH) 245.2 and277.8 nm (ε22270 and 3270); ν (CHBr₃) 3480 (OH), 1738 (acetate) and 1712cm⁻¹ (lactone); δ(CDCl₃) includes 0.92 (d,6 Hz,3 H), 0.97 (d,6 Hz,3H),1.06 (d,6 Hz,3H), 1.09 (d,6 Hz,3H), 1.77 (s,3H), 1.92 (d,15 Hz,1H), 2.17(s,3H), 3.09 (d,10 Hz, 1H), 3.16 (s,3H), 3.29 (d,15 Hz,1H), 3.32 (m,1H),3.84 (s,3H), 4.04 (d,6 Hz,1H), 5.54 (m,2H).

EXAMPLE 4 (13R)-(2'-Methoxyethoxymethoxy)-23(E)-methoxyimino FactorA,5-acetate

A solution of 2-methoxyethoxymethyl chloride (55 mg) in dichloromethane(250 μl) was added with stirring to a mixture of diisopropylethylamine(153 μl) and Intermediate 4 (100 mg) in dichloromethane (250 μl). Themixture was maintained at ca 20° for 5 days. Ether (50 ml) was added andthe mixture was washed with saturated sodium hydrogen carbonate andwater and dried. Removal of solvent gave a yellow foam which waspurified by medium pressure column chromatography on silica (80 g, MerckKieselgel 60, 230-400 mesh). Elution with dichloromethane:ether (6:1)gave the title compound as a white foam (70 mg); λmax (EtOH) 245.2 and277.4 nm (ε25330 and 2695); νmax (CHBr₃) 3540, 3420 (OH), 1736 (acetate)and 1712 cm⁻¹ (lactone); δ(CDCl₃) includes 0.92 (d,6 Hz,3H), 0.98 (d,6Hz,3H), 1.07 (d,6 Hz,3H), 1.10 (d,6 Hz,3H), 1.77 (s,3H), 1.91 (d, 15Hz,1H), 2.16 (s,3H), 3.29 (d,15 Hz,1H), 3.32 (m,1H), 3.39 (s,3H), 3.83(s,3H), 4.04 (d,6 Hz,1H), 4.5-4.8 (m,4H), 5.54 (m,2H).

EXAMPLE 5 13,23(E)-Bis(methoxyimino) Factor A, 5-acetate

A solution containing Intermediate 5 (56 mg), methoxyamine hydrochloride(43 mg) and anhydrous sodium acetate (41 mg) in methanol (10 ml) wasstood at 20° for 2 days then evaporated to near dryness. The resultantmixture was shaken with ethyl acetate and water and the organic phasewas washed successively with 0.5N hydrochloric acid and water. The driedorganic phase was evaporated and the crude product was purified bychromatography over Merck Keiselgel 60, 230-400 mesh (80 ml). Elution ofthe column with hexane:ethyl acetate (3:1) afforded the title compoundas a white foam (32 mg); [α]_(D) ²¹ +61° (c 1.11, CHCl₃); λmax (EtOH)247 nm (ε31,400), λmax (CHBr₃) (cm⁻¹) 3480 (OH), 1732 (OAc) 1712 (CO₂ R)δ(CDCl₃) include 5.54 (m;2H), 5.14 (m;1H), 3.84 (s;3H), 3.81 (s;3H),3.36 (m;1H), 3.29 (d15;1H), 3.14 (m;1H), 2.17 (s;3H), 1.91 (d15;1H),1.76 (s;3H), 1.66 (s;3H), 1.63 (s;3H), 1.21 (d6;3H), 1.06 (d6;3H), 0.96(d6;3H, 0.92 (d6;3H).

EXAMPLE 6 13,23(E)-Bis(methoxyimino) Factor A

A solution containing Example 5 (22 mg) and 1N sodium hydroxide (0.1 ml)in methanol (5 ml) was stirred in an ice bath for 1.3 h. The solutionwas diluted with ether (20 ml) and washed successively with 0.5Nhydrochloric acid and water. The dried organic phase was evaporated toafford the title compound as an off-white foam (12 mg); νmax (CHBr₃)(cm⁻¹) 3500 (OH), 1710 (C═O); δ(CDCl₃) include 5.18 (d9;1H), 5.12(m;1H), 4.30 (m;1H), 3.84 (s;3H), 3.81 (s;3H), 3.31 (m;1H), 3.28(d14;1H), 3.12 (m;1H), 1.88 (s;3H), 1.66 (s;3H), 1.63 (s;3H), 1.19(d6;3H), 1.05 (d6;3H), 0.96 (d6;3H), 0.91 (d6;3H).

EXAMPLE 7 (13S)-Methoxy- 23(E)-methoxyimino Factor A, 5-acetate

To a sample of Intermediate 6 (107 mg) under nitrogen was addedtrimethyloxonium tetrafluoroborate (230 mg),1,8-bisdimethylaminonaphthalene (330 mg) and dichloromethane (0.9 ml).The reaction mixture was left stirring at room temperature undernitrogen for 2 h, then the source of nitrogen removed. The reactionmixture was left stirring a further 72 h at room temperature then pouredinto water (30 ml) and extracted with diethyl ether (2×50 ml). Theorganic phase was washed with 2N hydrochloric acid, saturated sodiumbicarbonate solution, water and brine and then dried (MgSO₄). Removal ofsolvent gave a yellow foam (55 mg) which was combined with the crudeproduct (35 mg) from a similar reaction (using 47 mg of startingmaterial). The material was purified by medium pressure columnchromatography on silica (35 g, Merck Kieselgel 60, 230-400 mesh).Elution with dichloromethane:ethyl acetate (20:1) gave the titlecompound as a white foam (24 mg); δ(CDCl₃) includes 0.92 (d, 6 Hz,3H),0.96 (d,6 Hz,3H), 1.06 (d,6 Hz,3H), 1.12 (d,6 Hz,3H), 1.77 (s,3H), 2.17(s,3H), 3.28 (d,15 Hz,1H), 1.92 (d,15 Hz,1H), 3.32 (m,5H), 3.84 (s,3H),4.06 (d,6 Hz,1H), 5.53 (m,2H).

EXAMPLE 8 (13S)-(2'-Methoxyethoxymethoxy)-23(E)-methoxyimino Factor A,5-acetate

To a sample of Intermediate 6 (100 mg) was added a solution ofN,N-diisopropylethylamine (153 μl) in dichloromethane (250 μl) followedby a solution of 2-methoxyethoxymethyl chloride (55 mg) indichloromethane (250 μl). The reaction mixture was left stirring at roomtemperature for 140 h, then was poured into dichloromethane (50 ml) andwashed with 2N hydrochloric acid, saturated sodium bicarbonate solution,water and brine. The organic phase was dried (MgSO₄) and solvent removedto give a beige solid (111 mg) which was purified by medium pressurecolumn chromatography on silica (40 g, Merck Kieselgel 60, 230-400mesh). Elution with dichloromethane:ethyl acetate (10:1) gave the titlecompound as a white foam (58 mg); [α]_(D) ²¹ +77° (C=0.37, CH₂ Cl₂);λmax (EtOH) 245.0 nm ε27900 (E₁ ¹ 355); νmax 3540+3450 (OH) 1732(acetate) and 1710 cm⁻¹ (ester); δ(CDCl₃) includes 0.91 (d,6 Hz,3H),0.97 (d,6 Hz,3H), 1.06 (d,6 Hz,3H), 1.12 (d,6 Hz,3H), 1.76 (s,3H), 2.15(s,3H), 3.29 (d,14 Hz,1H), 3.32 (m,1H), 3.39 (s,3H), 3.82 (s,3H), 3.93(s,1H), 4.04 (d,6 Hz,1H), 4.67 (s,2H), 5.53 (m,2H).

The following are examples of formulations according to the invention.The term `Active Ingredient` as used hereinafter means a compound of theinvention.

MULTIDOSE PARENTERAL INJECTION EXAMPLE 1

    ______________________________________                                                        % w/v Range                                                   ______________________________________                                        Active ingredient  2.0    0.1-6.0% w/v                                        Benzyl alcohol     1.0                                                        Polysorbate 80    10.0                                                        Glycerol formal   50.0                                                        Water for Injections to                                                                         100.0                                                       ______________________________________                                    

Dissolve the active ingredient in the polysorbate 80 and glycerolformal. Add the benzyl alcohol and make up to volume with Water forInjections. Sterilize the product by conventional methods, for examplesterile filtration or by heating in an autoclave and packageaseptically.

EXAMPLE 2

    ______________________________________                                                       % w/v Range                                                    ______________________________________                                        Active ingredient                                                                              4.0     0.1-7.5% w/v                                         Benzyl alcohol   2.0                                                          Glyceryl triacetate                                                                            30.0                                                         Propylene glycol to                                                                            100.0                                                        ______________________________________                                    

Dissolve the active ingredient in the benzyl alcohol and glyceryltriacetate. Add the propylene glycol and make up to volume. Sterilizethe product by conventional pharmaceutical methods, for example sterilefiltration, and package aseptically.

EXAMPLE 3

    ______________________________________                                                        %           Range                                             ______________________________________                                        Active ingredient 2.0    w/v    0.1-7.5% w/v                                  Ethanol           36.0   v/v                                                  Non-ionic surfactant                                                                            10.0   w/v                                                  (e.g. Synperonic PE L44*)                                                     Propylene glycol to                                                                             100.0                                                       ______________________________________                                         *Trademark of ICI                                                        

Dissolve the active ingredient in the ethanol and surfactant and make upto volume. Sterilize the product by conventional pharmaceutical methods,for example sterile filtration, and package aseptically.

EXAMPLE 4

    ______________________________________                                                        %           Range                                             ______________________________________                                        Active Ingredient 2.0    w/v    0.1-3.0% w/v                                  Non-ionic surfactant                                                                            2.0    w/v                                                  (e.g. Synperonic PE F68*)                                                     Benzyl alcohol    1.0    w/v                                                  Miglyol 840**     16.0   v/v                                                  Water for Injections to                                                                         100.0                                                       ______________________________________                                         *Trademark of ICI                                                             **Trademark of Dynamit Nobel                                             

Dissolve the active ingredient in the Miglyol 840. Dissolve thenon-ionic surfactant and benzyl alcohol in most of the water. Preparethe emulsion by adding the oily solution to the aqueous solution whilehomogenising using conventional means. Make up to volume. Asepticallyprepare and package aseptically.

Aerosol spray

    ______________________________________                                                        % w/w  Range                                                  ______________________________________                                        Active Ingredient  0.1     0.01-2.0% w/w                                      Trichloroethane   29.9                                                        Trichlorofluoromethane                                                                          35.0                                                        Dichlorodifluoromethane                                                                         35.0                                                        ______________________________________                                    

Mix the Active Ingredient with trichloroethane and fill into the aerosolcontainer. Purge the headspace with the gaseous propellant and crimp thevalve into position. Fill the required weight of liquid propellant underpressure through the valve. Fit with actuators and dust-caps.

Tablet Method of manufacture--wet granulation

    ______________________________________                                                         mg                                                           ______________________________________                                        Active Ingredient  250.0                                                      Magnesium stearate 4.5                                                        Maize starch       22.5                                                       Sodium starch glycolate                                                                          9.0                                                        Sodium lauryl sulphate                                                                           4.5                                                        ______________________________________                                    

Microcrystalline cellulose to tablet core weight of 450 mg Addsufficient quantity of a 10% starch paste to the active ingredient toproduce a suitable wet mass for granulation. Prepare the granules anddry using a tray or fluid-bed drier. Sift through a sieve, add theremaining ingredients and compress into tablets.

If required, film coat the tablet cores using hydroxypropylmethylcellulose or other similar film-forming material using either an aqueousor non-aqueous solvent system. A plasticizer and suitable colour may beincluded in the film-coating solution.

Veterinary tablet for small/domestic animal use Method ofmanufacture--dry granulation

    ______________________________________                                                             mg                                                       ______________________________________                                        Active Ingredient      50.0                                                   Magnesium stearate      7.5                                                   Microcrystalline cellulose to tablet                                                                 75.0                                                   core weight of                                                                ______________________________________                                    

Blend the active ingredient with the magnesium stearate andmicrocrystallise cellulose. Compact the blend into slugs. Break down theslugs by passing through a rotary granulator to produce free-flowinggranules. Compress into tablets.

The tablet cores can then be film-coated, if desired, as describedabove.

Veterinary intrammary injection

    ______________________________________                                                         mg/dose      Range                                           ______________________________________                                        Active Ingredient          150 mg     0.05-1.0 g                              Polysorbate 60                                                                            3.0% w/w                                                          White Beeswax                                                                             6.0% w/w       to 3 g     to 3 or 15 g                            Arachis oil                                                                              91.0% w/w                                                          ______________________________________                                    

Heat the arachis oil, white beeswax and polysorbate 60° to 160° C. withstirring. Maintain at 160° C. for two hours and then cool to roomtemperature with stirring. Aseptically add the active ingredient to thevehicle and disperse using a high speed mixer. Refine by passing througha colloid mill. Aseptically fill the product into sterile plasticsyringes.

Veterinary slow-release bolus

    ______________________________________                                                   % w/w         Range                                                ______________________________________                                        Active Ingredient            0.25-2 g                                         Colloidal silicon                                                                          2.0                                                              dioxide                                                                       Microcrystalline             to required fill weight                          cellulose to 100.0                                                            ______________________________________                                    

Blend the active ingredient with the colloidal silicon dioxide andmicrocrystalline cellulose by using a suitable aliquot blendingtechnique to achieve a satisfactory distribution of active ingredientthroughout the carrier. Incorporate into the slow release device andgive (1) a constant release of active ingredient or (2) a pulsed releaseof active ingredient.

Veterinary oral drench

    ______________________________________                                                     % w/v    Range                                                   ______________________________________                                        Active Ingredient                                                                             0.35      0.01-2% w/v                                         Polysorbate 85 5.0                                                            Benzyl alcohol 3.0                                                            Propylene glycol                                                                             30.0                                                           Phosphate buffer                                                                             as pH 6.0-6.5                                                  Water to       100.0                                                          ______________________________________                                    

Dissolve the active ingredient in the Polysorbate 85, benzyl alcohol andthe propylene glycol. Add a proportion of the water and adjust the pH to6.0-6.5 with phosphate buffer, if necessary. Make up to final volumewith the water. Fill the product into the drench container.

Veterinary oral paste

    ______________________________________                                                        % w/w  Range                                                  ______________________________________                                        Active Ingredient 4.0      1-20% w/w                                          Saccharin sodium  2.5                                                         Polysorbate 85    3.0                                                         Aluminium distearate                                                                            5.0                                                         Fractionated coconut oil to                                                                     100.0                                                       ______________________________________                                    

Disperse the aluminium distearate in the fractionated coconut oil andpolysorbate 85 by heating. Cool to room temperature and disperse thesaccharin sodium in the oily vehicle. Disperse the active ingredient inthe base. Fill into plastic syringes.

Granules for veterinary in-feed administration

    ______________________________________                                                          % w/w  Range                                                ______________________________________                                        Active Ingredient    2.5     0.05-5% w/w                                      Calcium sulphate, hemi-hydrate to                                                                 100.0                                                     ______________________________________                                    

Blend the Active Ingredient with the calcium sulphate. Prepare thegranules using a wet granulation process. Dry using a tray or fluid-beddrier. Fill into the appropriate container.

Veterinary Pour-on

    ______________________________________                                                            % w/v Range                                               ______________________________________                                        Active Ingredient      2.0    0.1 to 30%                                      Demethyl sulphoxide   10.0                                                    Methyl Isobutyl ketone                                                                              30.0                                                    Propylene glycol (and pigment) to                                                                   100.0                                                   ______________________________________                                    

Dissolve the active ingredient in the dimethyl sulphoxide and the methylisobutyl ketone. Add the pigment and make up to volume with thepropylene glycol. Fill into the pour-on container.

Emulsifiable Concentrate

    ______________________________________                                        Active ingredient         50    g                                             Anionic emulsifier        40    g                                             (e.g. Phenyl sulphonate CALX)                                                 Non-ionic emulsifier      60    g                                             (e.g. SYNPERONIC NP13)*                                                       Aromatic solvent (e.g. Solvesso 100) to                                                                 1     liter.                                        ______________________________________                                         *Trademark of ICI                                                        

Mix all ingredients, stir until dissolved.

Granules

    ______________________________________                                        (a)    Active ingredient      50    g                                                Wood resin             40    g                                                Gypsum granules (20-60 mesh) to                                                                      1     kg                                               (e.g. Agsorb 100A)                                                     (b)    Active ingredient      50    g                                                SYNPERONIC NP13*       40    g                                                Gypsum granules (20-60 mesh) to                                                                      1     kg.                                       ______________________________________                                         *Trademark of ICI                                                        

Dissolve all ingredients in a volatile solvent e.g. methylene chloride,add to granules tumbling in mixer. Dry to remove solvent.

We claim:
 1. Compounds of formula (1) ##STR15## wherein R¹ represents amethyl, ethyl or isopropyl group; Y¹ is --CH₂ --, Y² is --CH-- and Xrepresents ##STR16## where R² and R³ together with the carbon atom towhich they are attached represents >C═NOR⁷, where R⁷ represents ahydrogen atom, a C₁₋₈ alkyl group or a C₃₋₈ alkenyl group and the group>C═NOR⁷ is the E configuration, or --Y¹ --X--Y² represents --CH₂--CH═C--;R⁴ represents an hydroxy, methoxy or acyloxy group and R⁵represents a hydrogen atom or R⁴ and R⁵ together with the carbon atomsto which they are attached represents >C═O or >C═NOR^(7a), where R^(7a)is as defined above for R⁷ ; and one of R⁸ and R⁹ represents C₁ -C₆alkoxy group optionally interrupted by an oxygen atom or a C₁ -C₆ alkoxygroup and the other represents a hydrogen atom or R⁸ and R⁹ togetherwith the carbon atom to which they are attached represent >C═NOR^(7b),where R^(7b) is as defined above for R⁷, and salts thereof.
 2. Compoundsaccording to claim 1 in which R¹ is an isopropyl group.
 3. Compoundsaccording to claim 1 in which R⁸ or R⁹ is a methoxy group or --OCH₂ OCH₂CH₂ OCH₃.
 4. Compounds according to claim 1 in which R¹ is an isopropylgroup, Y¹ is --CH₂ --, Y² is --CH--, X is --C(R²)(R³)--, in which R² andR³ together with the carbon atom to which they are attached represent>C═CH₂ or >C═NOCH₃ (where the group >C═NOCH₃ is in the E configuration),R⁴ is a hydroxy or acetoxy group and R⁵ is a hydrogen atom; and one ofR⁸ and R⁹ represents a methoxy group or a group --OCH₂ OCH₂ CH₂ OCH₃ andthe other represents a hydrogen atom or R⁸ and R⁹ together with thecarbon atom to which they are attached represent >C═NOCH₃.
 5. Compoundsaccording to claim 1 in whichR¹ is an isopropyl group, Y¹ is --CH₂ --,Y² is --CH--, X represents >C═NOCH₃ --, R⁴ is an acetoxy group, R⁵ is ahydrogen atom, R⁸ is a hydrogen atom and R⁹ is a methoxy group; R¹ is anisopropyl group, Y¹ is --CH₂ --, Y² is --CH--, X represents >C═NOCH₃, R⁴is an acetoxy group, R⁵ is a hydrogen atom, R⁸ is a methoxy group and R⁹is a hydrogen atom; and R¹ is an isopropyl group, Y¹ is --CH₂ --, Y² is--CH--, X represents >C═NOCH₃, R⁴ is an acetoxy group, R⁵ is a hydrogenatom, R⁸ is a group --OCH₂ OCH₂ CH₂ OCH₃ and R⁹ is a hydrogen atom.
 6. Apharmaceutical composition containing a pesticidally effective amount ofat least one compound as claimed in claim 1 together with apharmaceutically acceptable carrier.
 7. A veterinary compositioncontaining a pesticidally effective amount of at least one compound asclaimed in claim 1 and a veterinary acceptable carrier.
 8. A pesticidalcomposition containing a pesticidally effective amount of a compound asclaimed in claim 1 and a pesticidally acceptable carrier.
 9. A method ofcontrolling insect, acarine or nematode pests which comprises applyingan amount of a compound according to claim 1 effective in combattingpests to the pests or a locus of said pests.